Composition that inhibits or destroys at least one unicellular living organism containing fluorine F- and Lithium li+

ABSTRACT

The invention is directed to a composition and a method for using the same which composition inhibits or destroys unicellular living organisms. The composition is particularly effective in combating sexually transmitted diseases and as an antiseptic agent. The composition of the invention comprises lithium and ionic or ionizable, fluorine, for example, lithium fluoride. A suitable excipient such as KATHON may also be present.

This application is a continuation of application Ser. No. 08/125,526, filed Sep. 22, 1993, which is a continuation of pending prior application Ser. No. 07/691,472, as originally filed on Apr. 25, 1991, now abandoned which is a continuation-in-part of application Ser. No. 07/246,982, filed Sep. 20, 1988; now U.S. Pat. No. 5,063,064.

BACKGROUND OF THE INVENTION

The invention relates to the inhibition or destruction of unicellular living organisms such as protozoa, microbes, bacteria, gametes, fungi, yeasts or the like, and viruses. It hence relates especially to the technical fields of local contraception, of antibiotic therapy and of antisepsis, whether in the context of pharmacy or of cosmetics, as well as to that of disinfection.

Many substances and compositions that inhibit or destroy unicellular living organisms are already known. Unpublished European Patent Application No. 86/402,716.4, filed on 8 Dec., 1986 and U.S. patent application Ser. No. 07/053,374 filed on 22 May, 1986 by the applicant, mention such substances and compositions, and already teach that it is advantageous to use a compound that liberates F⁻ ions alone or in combination with another primary active principle.

Moreover, lithium in the ionic state is already known in various therapeutic applications: in neurology, by way of a normothymic agent, in rhumatology or urology, by way of a uric acid-eliminating gent, in the field of dental care, by way of antiseptic PYOREX, registered trademark), and in the treatment of conditions of the airways in pneumology (THIOPHEOL).

Lithium fluoride is also already known as a chemical compound (The MERCK INDEX, TENTH EDITION, page 793, reference 5357). This compound is not, however, used by way of a medicinal product, or for inhibiting or destroying unicellular living organisms.

European Patent Application 0 055 109 describes an anti-caries composition comprising a fluorine salt and a carbohydrate. Among fluorides used by way of an anti-caries agent, lithium fluoride is mentioned. However, this document does not draw attention to any particular property due to the lithium fluoride for inhibiting or destroying microorganisms. In effect, in the combating of dental caries, fluorine has a known action, which does not consist in inhibiting or destroying microorganisms.

European Patent Application 0,162,574 describes a composition for oral/dental hygiene, such as a dentifrice, comprising a fluorine salt, a zinc salt, a buffering agent and an excipient. Among the fluorine salts referred to, lithium fluoride is mentioned. However, the fluorine salts are used in the context of that document only by way of anti-caries agent, as is taught in European Patent Application 0,055,109. Moreover, European Patent Application 0,162,574 also mentions the possibility of using a cationic surfactant agent by way of an antiseptic and antibacterial. However, this document does not describe a composition that makes it possible to inhibit or destroy microorganisms containing lithium fluoride.

The objects of the invention are to remedy the known drawbacks of the prior art, namely:

to reduce the necessary concentrations of active principles and/or activating principles while retaining the same efficacy, in order to limit or avoid side effects,

to propose a composition that inhibits or destroys unicellular living organisms and which is safely administrable systemically or parenterally,

to provide a new compound which is usable by way of a medicinal product or bactericidal, antibiotic, virucidal, antiseptic or disinfectant product or contraceptive,

to propose other therapeutic applications of ionic lithium, and

to provide a composition which is active against certain pathogenic organisms against which no remedy is known.

THE SUMMARY OF THE INVENTION

The invention hence proposes a spermicidal composition, a composition enabling sexually transmitted diseases to be combated, an antiseptic composition for the local disinfection of the human body--in particular of the skin, the mucosae, the limbs, etc--a composition for the disinfection of surfaces such as floors or instruments, and an antibiotic composition, and more generally a composition intended for destroying or inhibiting unicellular living organisms such as protozoa, microbes, bacteria, gametes, fungi, yeasts, viruses or the like, in therapeutic, contraceptive, sanitary or agricultural applications, characterized in that they contain:

ionic or ionisable lithium, or

ionic or ionisable lithium and ionic or ionisable fluorine, in particular lithium fluoride, or

A) lithium fluoride and B) a suitable excipient, or

A) lithium fluoride, B) an active principle suitable for the application in question--in particular a surfactant detergent such as a quaternary ammonium compound, for example benzalkonium chloride, or a nonoxinol, or an antibiotic or a bactericide, or a sporicide, or a fungicide, or a virucide, or an antiseptic, or a disinfectant, or a spermicide--and C) a suitable excipient, or

A) lithium fluoride, B) an active principle suitable for the application in question--in particular as referred to above--, C) a preservative agent such as KATHON CG (registered trademark) and D) a suitable excipient.

A composition according to the invention can also contain in addition, another fluorine salt, in particular sodium fluoride.

The invention also proposes a pharmaceutical composition containing ionic and ionizable fluorine and ionic or ionisable lithium, in particular lithium fluoride. The invention also proposes such a composition containing in addition, an active principle suitable for the therapeutic application in question--in particular a surfactant detergent such as quaternary ammonium compound, for example benzalkonium chloride or a nonoxinol, or an antibiotic, or a bactericide, or a spermicide, or a fungicide, or a virucide. The invention also proposes such a composition containing, in addition, an active principle with the exception of a zinc salt.

The invention also relates to the application of lithium Li⁺ cations for manufacturing a product intended for destroying or inhibiting gametes--in particular spermatozoa--by way of a local contraceptive.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions and Li⁺ ions--in particular lithium fluoride--for manufacturing a composition intended for combating sexually transmitted diseases.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions and Li⁺ ions--in particular lithium fluoride--for manufacturing an antibiotic composition.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions and Li⁺ ions--in particular lithium fluoride --for manufacturing an antiseptic or disinfectant composition.

The invention also relates to the application at least one compound capable of liberating F⁻ ions, at least one compound capable of liberating Li⁺ ions--in particular lithium fluoride--and at least one active principle--in particular a surfactant detergent such as a quaternary ammonium compound, for example benzalkonium chloride, or a nonoxinol--for manufacturing a spermicidal composition.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions, at least one compound capable of liberating Li⁺ ions--in particular lithium fluoride--and at least one active principle--in particular a surfactant detergent such as a quaternary ammonium compound, for example benzalkonium chloride, or a nonoxinol--for manufacturing a compound intended for combating sexually transmitted diseases.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions, at least one compound capable of liberating Li⁺ ions--in particular lithium fluoride--and at least one active principle--in particular a surfactant detergent such as a quaternary ammonium compound, for example benzalkonium chloride, or a nonoxinol--for manufacturing an antiseptic or disinfectant composition.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions, at least one compound capable of liberating Li⁺ ions--in particular lithium fluoride--and at least one antibiotic principle for manufacturing an antibiotic product containing sufficiently low concentrations of F⁻, Li⁺ and antibiotic principle to enable it to be administered orally or parenterally.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions, at least one compound capable of liberating Li⁺ ions--in particular lithium fluoride--at least one virucidal principle--in particular a surfactant detergent such as a quaternary ammonium compound, for example benzalkonium chloride--for manufacturing a virucidal product containing sufficiently low concentrations of F⁻, Li⁺ and primary virucidal principle to enable it to be administered orally or parenterally.

The invention also relates to the application of at least one compound capable of liberating F⁻ ions, at least one compound capable of liberating Li⁺ ions--in particular lithium fluoride--and at least one bactericidal or antibiotic or fungicidal principle, for manufacturing a composition intended for combating pathogenic microorganisms such as chlamydiae, Gardnerella vaginalis, Ducrey's bacillus, Candida albicans, Aspergillus, Streptococcus, Proteus vulgaris, Haemophilus influenzae, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus, Mycobacterium, Neisseria gonorrhoeae, Irichomonas, Treponema, Sarcina lutea, Bacillus subtilis, Klebsiella pneumoniae and Enterococcus.

The invention hence demonstrates, on the one hand, the surprising synergistic effect of the combination of F⁻ anions with Li⁺ cations for destroying or inhibiting unicellular living organisms, and by way of a medicinal product, contraceptive, cosmetic, bactericide, antibiotic, virucide or disinfectant, or the other hand, new applications given to Li⁺ ions, and finally a surprising activity of the combination of F⁻, Li⁺ and a primary active principle, enabling the concentrations to be reduced considerably and, in particular, products that are usable orally or parenterally to be obtained.

The invention also consists in having selected lithium fluoride among the possible fluorides mentioned in the prior art, and in having qualitatively and quantitatively demonstrated unexpected properties of lithium fluoride.

EXAMPLES AND PREFERRED EMBODIMENTS

The invention is illustrated by the examples below; the terminology and the methodologies used below have already been defined in unpublished European Patent Application No. 86/402,716.4, filed on 8 Dec., 1986 and U.S. patent application Ser. No. 07/053,374 filed on 22 May, 1987 by the applicant, and are integrated in the present specification and will hence not be fully specified again.

A benzalkonium chloride solution used in the trials contained a minimum of 90% by weight of C14 benzalkonium chloride of formula:

(C₆ H₅ --CH₂ --CH₃ NCH₃ --C₁₄ H₂₉ +, Cl⁻) or myristyldimethylbenzylammonium chloride. It is also possible to use cocodimethylbenzylammonium chloride or alkyldimethylbenzylammonium chloride, or another quaternary ammonium chloride. All the percentages given are percentages by weight.

KATHON⁻ CG (registered trademark) is a known preservative marketed by ROHM AND HAAS COMPANY (U.S.A.), and consisting of a mixture of two isothiazolines identified according to IUPAC nomenclature as 5-chloro 2-methyl-4-isothiazolin-3-one and 2-methyl-4 isothiazolin-3-one.

I--APPLICATION OF THE INVENTION IN THE FIELD OF LOCAL CONTRACEPTION Example 1

European Patent Applicaton No. 86/402,716.4 showed that F⁻ ions alone have a MIC (minimal inhibitory concentration), according to the SANDERS-CRAMER total spermicidal test according to IPPF standards of 5 ppm, equivalent to 0.0005%.

The same test carried out with lithium fluoride Lif alone showed that lithium fluoride alone has a MIC of the order of 3 ppm, equivalent to 0.0003%.

The same test carried out with lithium chloride LiCl alone showed that lithium chloride alone has a MIC of 20 ppm, equivalent to 0.002%.

A spermicidal effect of Li⁺ ions is hence observed. However, above all, the combination F⁻ /Li⁺ achieves an unexpected synergistic effect.

The proportions of lithium fluoride contained in a spermicidal composition according to the invention must be such that the titre of lithium fluoride which it can liberate in vivo is greater than 2.5 mg/l, in order to be effective without reaching doses that cause side reactions.

The proportions liberated in vivo depend, of course, on the galenical form used. For example, the following galenical forms yield good results: cream, jelly, pessary, pad, soluble sheet, tablet, foam. For example:

    ______________________________________                                         GALENICAL              LiF                                                     FORM                   (% by weight)                                           ______________________________________                                         CREAM                  0.55                                                    ______________________________________                                    

Example 2

Furthermore, European Patent Application No. 86/402,716.4 showed that the addition of 1 millilitre of solution containing 0.0001% of F⁻ anions (for example in the form of sodium fluoride) to 1 millilitre of composition containing benzalkonium chloride as primary active principle, in the context of the total spermicidal test according to IPPF standards, enables the MIC of the benzalkonium chloride to be lowered from a value of 0.006% (in the initial 1 millilitre) to 0.002% (in the initial 1 millilitre).

The same test carried out (in vitro) replacing the solution of 1 millilitre containing 0.0001% of F⁻ anions by a solution of 1 millilitre containing 0.0001% of lithium fluoride enabled as MIC of benzalkonium chloride of 0.0009% (in the initial 1 millilitre) to be obtained.

A further boosting effect is hence observed. The maximal results (in vitro) were obtained with a solution of lithium fluoride at a concentration of 0.0001%.

The following galenical forms may be used: cream, jelly, pessary, pad, soluble sheet, tablet, foam. For example:

CREAM:

    ______________________________________                                         benzalkonium chloride                                                                           0.90%                                                         LiF              0.55%                                                         ______________________________________                                    

excipients: emulsive agent, preservative such as KATHON (registered trademark), purified water, perfume qs. 100%.

JELLY: same formulations as the cream excipients: soluble derivatives of cellulose, glycerin, preservative such as KATHON (registered trademark), purified water, perfume qs. 100%.

PAD: impregnated with the cream defined above

TABLETS:

    ______________________________________                                         benzalkonium chloride                                                                           0.020 g                                                       LiF              0.010 g                                                       ______________________________________                                    

excipients: sodium bicarbonate, citric acid, colloidal silica, cellulose, magnesium stearate, lactose qs. 1 tablet.

SOLUBLE SHEET: same formulations as the cream excipients: polyvinilic alcohol, glycerin, preservative such as KATHON (registered trademark), purified water qs. 100%.

PESSARIES:

    ______________________________________                                         benzalkonium chloride                                                                           0.017 g                                                       LiF              0.010 g                                                       ______________________________________                                    

excipients: semi-synthetic glycerides, preservative such as KATHON (registered trademark) qs. 1 pessary.

A local contraceptive, in particular spermicidal, product according to the invention is characterized in that it contains between 0.20% and 0.75%--in particular, of the order of 0.55%--by weight of lithium fluoride and between 0.10% and 1.20% of benzalkonium chloride.

II--APPLICATION OF THE INVENTION FOR COMBATING THE MICROORGANISMS RESPONSIBLE FOR INFECTIONS SUCH AS SEXUALLY TRANSMITTED DISEASES Example 3

Combating Chlamydiae

The methodology of the tests performed in order to demonstrate the application of the invention in the context of combating chlamydiae is that described by Professors F. CATALAN, P. SEDNAOUI, A. MILOVANOVIC et al., Institut A. FOURNIER, PARIS, already mentioned in unpublished European Patent Application No. 86/402,716.4

The test was performed on 12 strains of Chlamydia trachomatis originating from hospital sources, in particular from cases of urethritis. All the trials were performed in duplicate, and the results compared with those for a listed control strain.

It was first verified that lithium fluoride at a concentration of 1 mg/l does not have toxic activity with respect to MAC COY cells. In addition, since the threshold of cytotoxic activity of benzalkonium chloride is 0.01% with respect to MAC COY cells, benzalkonium chloride is not toxic either on MAC COY cells under the conditions of the trials carried out.

The MIC corresponds to the concentration of benzalkonium chloride at and above which the average number of colonies in Petri dishes containing the active substance is less than one tenth of the average number of colonies in Petri dishes without active substance.

The MIC of benzalkonium chloride alone varied with respect to the 12 wild-type strains tested, from 12 to 180 mg/l. The MIC of benzalkonium chloride with respect to the listed test strain is 18 mg/l.

The MIC of benzalkonium chloride to which 1 mg/ml of lithium fluoride has been added varied with respect to these same 12 strains tested, from 8 to 160 mg/l. The MIC of benzalkonium chloride to which 1 mg/ml has been added with respect to the listed test strain is 13 mg/l.

A big improvement of the results with lithium fluoride is hence observed.

Example 4

Combating Gardnerella vaginalis

Two trials were carried out in order to illustrate the application of the invention in combating Gardnerella vaginalis.

In the first trial, the sample of bactericidal substance was incorporated directly in the specific culture edium of the microorganism. A concentration series of the bactericidal substance was prepared in the geometric ratio of 2, from 400 μg/ml to 1.56 μg/ml. The cell concentration used in the seeding of the Petri dishes was 10⁻³ per ml. A 24-hour incubation was performed at 37° C. and then, for each wild-type strain tested, the concentration of primary principle destroying all the microorganisms (see method of Professor F. CATALAN et al., Institut A. FOURNIER, PARIS) was determined. The trail was performed on 32 wild-type strains of Gardnerella vaginalis originating from hospital sources and with benzalkonium chloride and then nonoxinol 9 as active principles, and with sodium fluoride NaF at a concentration of 1 mg/l and then lithium fluoride LiF at a concentration of 1 mg/l as activating principles. The results obtained are correlated in the table below, which expresses the number of strains inhibited out of the 32 tested, in terms of the concentrations of primary active principle (benzalkonium chloride or nonoxinol).

    __________________________________________________________________________                Concentration in mg/l of primary                                               active principle (benzalkonium                                                 chloride or nonoxinol)                                              SUBSTANCE  1.56                                                                              3.125                                                                             6.25                                                                              12.5                                                                              25 50 100                                                                               200                                                                               400                                         __________________________________________________________________________     Benzalkonium chloride                                                                     0  0  1  28 31 32 -- -- --                                          Benzalkonium chloride +                                                                   0  0  7  30 32 -- -- -- --                                          NaF (1 mg/l)                                                                   Benzalkonium chloride +                                                                   0  0  13 32 -- -- -- -- --                                          LiF (1 mg/l)                                                                   Nonoxinol 9                                                                               0  0  0  0  4  6  10 25 32                                          Nonoxinol 9 +                                                                             0  0  0  1  12 20 26 31 32                                          NaF (1 mg/l)                                                                   Nonoxinol 9 +                                                                             0  0  0  1  16 25 30 32 --                                          LiF (1 mg/l)                                                                   __________________________________________________________________________

This table illustrates the surprising boosting effect of lithium fluoride compared with sodium fluoride. In effect, at equal concentrations, the number of strains inhibited in the presence of LiF is always markedly greater than the number of strains inhibited in the presence of NaF. The totally lethal concentration for benzalkonium chloride falls from 50 mg/l when it is alone, to 25 mg/l in the presence of NaF, to 12.5 mg/l in the presence of LiF.

The second trial carried out was performed according to the contact methodology described in AFNOR standard I 72-151, by counting the strains surviving after a 15-min active substance/microbacterial solution contact followed by a filtration on a membrane (porosity 0.22 μm).

In this trial, 26 wild-type strains of Gardnerella vaginalis were tested, originating from hospital sources. The starting solution contained 0.1% of benzalkonium chloride, and was then diluted to 10⁻¹, 10⁻², 10⁻³ and 10⁻⁴. The results are summarized in the table below, which gives the total number of strains surviving (out of the 26 tested) in terms of the dilution.

    ______________________________________                                         Dilution of the                                                                         Benzalkonium                                                                               Benzalkonium                                                                              Benzalkonium                                   0.1% strength                                                                           chloride    chloride + NaF                                                                            chloride + LiF                                 solution alone       (1 mg/l)   (1 mg/l)                                       ______________________________________                                         10.sup.-1                                                                               0           0          0                                              10.sup.-2                                                                               18          7          0                                              10.sup.-3                                                                               22          15         15                                             10.sup.-4                                                                               26          26         26                                             ______________________________________                                    

EXAMPLE 5

Combating Trichomonas vaginalis

The same methodology as in example 4 was used on 15 strains of Trichomonas vaginalis. The results are given in the following table, which indicates the number of strains inhibited out of the 15 tested, in terms of the concentrations of benzalkonium chloride:

    __________________________________________________________________________            Concentrations in mg/l of benzalkonium                                         chloride                                                                SUBSTANCE                                                                             1.56                                                                               3.125                                                                             6.25                                                                              12.5                                                                              25  50 100                                                                               200                                                                               400                                           __________________________________________________________________________     Benzalkonium                                                                          1   3   9 13 15  -- -- -- --                                            chloride                                                                       Benzalkonium                                                                          3   8  15 -- --  -- -- -- --                                            chloride + Lif                                                                 (1 mg/l)                                                                       __________________________________________________________________________

The MIC hence falls from 25 mg/l for benzalkonium chloride alone to 6.25 mg/l in the presence of LiF.

Example 6

The MIC of benzalkonium chloride, alone and in the presence of LiF at a concentration of 1 mg/l, was determined with respect to other wild-type strains originating from hospital sources namely Streptococcus (6 strains). Staphylococcus aureus (8 strains). Salmonella typhimurium (3 strains) and Sarcina lutea (1 strain).

The MIC values obtained are summarized below.

    ______________________________________                                                                      Benzalkonium chloride                                         Benzalkonium     in the presence of                                STRAIN      chloride alone   LiF at 1 mg/l                                     ______________________________________                                         Streptoccocus                                                                              18-20   mg/l     10     mg/l                                       Staphylococcus                                                                             1.56    mg/l     0.30   mg/l                                       aureus                                                                         Salmonella  17      mg/l     12     mg/l                                       typhimurium                                                                    Sarcina     15      mg/l     11     mg/l                                       lutea                                                                          ______________________________________                                    

EXAMPLE 7

Combating Ducrey's Bacillus

The methodology employed is that described by Professor F. CATALAN, P. SEDNAOUI, A. MILOVANOVIC et al., Institut A. FOURNIER, PARIS.

The trials were performed on 8 wild-type strains of DUCREY's Haemophilus. The following results were recorded with respect to these strains:

    ______________________________________                                                  MIC of the primary active principle with                              Primary  respect to DUCREY's bacillus (mg/l)                                   active   Primary active                                                                             in the presence                                                                           in the presence                                principle                                                                               principle alone                                                                            of NaF (1 mg/l)                                                                           of LiF (1 mg/l)                                ______________________________________                                         Benzalkonium                                                                            100         85         65                                             chloride                                                                       Nonoxinol 9                                                                             150         132.5      127.5                                          ______________________________________                                    

Lithium fluoride hence proves to be a markedly better activating principle than sodium fluoride.

EXAMPLE 8

Combating Candida albicans

With the same methodology, trials were performed on hospital strains of Candida albicans:

    ______________________________________                                                  MIC of the primary active table                                                                    in the                                                                         presence of LiF                                                                at 0.96 mg/l and                                                      in the   of KATHON CG                                      Primary  Primary    presence (registered trademark)                            active   active     of LiF at                                                                               preservative at                                   principle                                                                               principle  0.96 mg/l                                                                               0.234/mgl                                         ______________________________________                                         Benzalkronium                                                                           50 mg/l    135 mg/l 2 mg/l                                            chloride                                                                       ______________________________________                                    

EXAMPLE 9

Combating Aspergillus niger

With the same methodology, the following results were observed on hospital strains of Aspergillus niger:

    ______________________________________                                                  MIC of the primary active principle                                                                In the                                                                         presence of LiF                                                                at 0.96 mg/l and                                           Primary    In the   of KATHON CG                                      Primary  active     presence (registered trademark)                            active   principle  of LiF at                                                                               preservative at                                   principle                                                                               alone      0.96 mg/l                                                                               0.234 mg/l                                        ______________________________________                                         benzalkonium                                                                            50 mg/l    135 mg/l 100/mg/l                                          chloride                                                                       ______________________________________                                    

EXAMPLE 10

Combating Pyogenic Microorganisms

With the same methodology, the following results were observed on hospital strains of pyogenic microorganisms, namely 8-haemolytic streptococcus, Proteus vulgaris, Haemophilus influenzae:

    ______________________________________                                         Primary       MIC of the primary active principle                              active        β-Haemolytic                                                                         Proteus   Haemophilus                                 principle     streptococcus                                                                             vulgaris  influenzae                                  ______________________________________                                         Benzal-                                                                              Alone       50     mg/l  25   mg/l 200  mg/l                             konium                                                                               In the presence                                                                            25     mg/l  18.5 mg/l 150  mg/l                             chloride                                                                             of LiF at 1 mg/l                                                               and of KATHON                                                                  CG.sup.R preservative                                                          at 0.4 mg/l                                                              ______________________________________                                    

The preferred galenical forms which can, for example, be used for combating STD's are the same as in the application by way of a spermicide.

Nonoxinol 9 may also be used as the active principle, for example:

PESSARY:

    ______________________________________                                                nonoxinol    0.060   mg/l                                                      LiF          0.01    g                                                  ______________________________________                                    

excipients: semi-synthetic glycerides, preservative such as KATHON (registered trademark) qs. 1 pessary.

III--APPLICATION OF THE INVENTION IN THE FIELD OF ANTI-SEPSIS AND DISINFECTION Example 11

French standard NF T 72-150, March 1981, was followed by the trials carried out. The trials were performed on the strains defined by the AFNOR standard.

The results obtained on the strains defined by the AFNOR standard for benzalkonium chloride alone and for benzalkonium chloride with the addition of 1 mg/l of sodium fluoride were already given in unpublished European Patent Application 86/402,716.4, and have been recorded again.

These results are as follows:

    ______________________________________                                                                  MINIMAL INHIBITORY                                                             CONCENTRATION (MIC)                                            MINIMAL INHIBITORY                                                                             OF BENZALKONIUM                                                CONCENTRATION (MIC)                                                                            CHLORIDE IN THE                                                OF BENZALKONIUM PRESENCE OF                                           STRAIN   CHLORIDE ALONE  NaF AT 1 mg/l                                         ______________________________________                                         Pseudomonas                                                                             31.25     mg/l      18      mg/l                                      aeruginosa                                                                     CNCM A 22                                                                      Escherichia                                                                             6.57      mg/l      3       mg/l                                      coli                                                                           CNCM 54 127                                                                    Staphylococcus                                                                          1.56      mg/l      1.1     mg/l                                      aureus                                                                         Oxford strain                                                                  CNCM 53 154                                                                    Streptococcus                                                                           4         mg/l      3.6     mg/l                                      faecalis                                                                       CNCM 5 855                                                                     Mycobacterium                                                                           30        mg/l      26      mg/l                                      smegmatis                                                                      CNCM7 326                                                                      ______________________________________                                    

The MIC obtained for benzalkonium chloride to which lithium fluoride was added at a concentration of 1 mg/l is as follows:

    ______________________________________                                                        MINIMAL INHIBITORY CONCENTRA-                                                  TION (MIC) OF BENZALKONIUM                                                     CHLORIDE IN THE PRESENCE OF                                     STRAIN         LiF AT 1 mg/l                                                   ______________________________________                                         Pseudomonas aeruginosa                                                                        15           mg/liter                                           CNCM A 22                                                                      Escherichia Coli                                                                              2            mg/liter                                           CNCM 54 127                                                                    Staphylococcus faecalis                                                                       1.1          mg/liter                                           Oxford strain CNCM 53 154                                                      Streptococcus faecalis                                                                        2.8          mg/liter                                           CNCM 5 855                                                                     Mycobacterium smegmatis                                                                       22           mg/liter                                           CNCM 7 326                                                                     ______________________________________                                    

Example 12

The same trials as in example 11 were carried out on wild-type strains originating from hospital sources, namely 120 strains of Pseudomonas, 200 strains of Escherichia coli, 300 strains of Staphylococcus, 200 strains of Streptococcus and 50 strains of Mycobacterium.

The results obtained on the wild-type strains are summarized in the following table:

    ______________________________________                                                                         MIC of                                                        MIC of           benzalkonium                                                  benzalkonium     chloride in the                                               chloride         presence of                                    STRAIN         alone            LiF (1 mg/l)                                   ______________________________________                                         Pseudomonas aeruginosa                                                                        30 to 120                                                                               mg/l    15 to 80                                                                              mg/l                                    Escherichia coli                                                                              6 to 25  mg/l    1.8 to 16                                                                             mg/l                                    Staphylococcus aureus                                                                         2 to 13  mg/l    0.8 to 8                                                                              mg/l                                    Streptococcus faecalis                                                                        5 to 30  mg/l    2.5 to 22                                                                             mg/l                                    Mycobacterium smegmatis                                                                       32 to 120                                                                               mg/l    20 to 90                                                                              mg/l                                    ______________________________________                                    

Example 13

The same trials as in example 12 were carried out on hospital strains of various microorganisms. The results obtained are as follows:

    ______________________________________                                                     MIC of     MIC of benzalkonium                                                 benzalkonium                                                                              chloride in mg/l in the                                 STRAIN      alone in mg/l                                                                             presence of Lif at 1 ppm                                ______________________________________                                         Neisseria   1.15       0.60                                                    gonorrhoea                                                                     Trichomonas 1.30       0.90                                                    vaginalis                                                                      Candida     50         35                                                      albicans                                                                       Gardenerella                                                                               50         41                                                      vaginalis                                                                      Ducrey's    75         62                                                      bacillus                                                                       Streptococcus                                                                              15         9                                                       faecalis                                                                       Staphylococcus                                                                             1.56       1                                                       aureus                                                                         Aspergillus 85         80                                                      niger                                                                          Escherichia 8          6.5                                                     coli                                                                           Sarcina lutea                                                                              11         7.5                                                     Bacillus    8          5.4                                                     subtilis                                                                       ______________________________________                                    

Example 14

Trials similar to those carried out in Examples 12 and 13 were carried out on 80 hospital strains of multi-resistant Escherichia coli producing plasmid β-lactamase. The results are as follows:

    ______________________________________                                                 MIC of the primary active principle in mg/l                                                             in the presence                                                                of LiF at 0.96                                                                 mg/l and of                                                                    KATHON CG                                               Primary                (registered                                   primary   active                 trademark) pre-                               active    principle in the presence                                                                             servative at                                  principle alone     LiF at 0.96 mg/l                                                                            0.47 mg/l                                     ______________________________________                                         Benzalkonium                                                                             8 to 12   6 to 9       4 to 6                                        cloride                                                                        ______________________________________                                    

Example 15

The bactericidal activity of seven commercial local disinfectant antiseptics was compared with that of a foaming paste according to the invention, containing:

    ______________________________________                                         benzalkonium chloride    2%                                                    KATHON CG (registered trademark)                                                                        0.1%                                                  lithium fluoride         0.004%                                                sodium fluoride          1.5%                                                  excipient: foaming base  qs.                                                   and purified water                                                             ______________________________________                                    

The methodology is that defined by AFNOR standard I 72-150, using the products under the conditions specified by the manufacturer as regards the concentrations and conditions of use.

In the table below, the sign + indicates that a growth of the microorganisms was observed, the sign ++ indicates that a very strong growth of the microorganisms was observed, and the sign - indicates that no growth of the microorganisms was observed.

Septivon-Lavril is a foaming solution marketed by Laboratoires CLIN MIDY (Paris, France).

Solubacter is a registered trademark denoting a foaming solution marketed by Laboratoires INNOTHERA (Arcueil, France).

Cyteal is a foaming solution marketed by Laboratoires SINBIO (Paris, France).

Hibiscrub is a foaming solution marketed by I.C.I. PHARMA (Cergy, France).

Hibisprint is an alcoholic solution marketed by I.C.I.--PHARMA (Cergy, France).

Hexomedine is a registered trademark denoting a non foaming solution marketed by THERAPLIX S.A. (France).

Cetavlon is an antiseptic solution marketed by I.C.I.--PHARMA (Cergy, France).

The results obtained on Escherichia coli, Staphylococcus aureus, Streptococcus group A and Group C streptococcus are as follows:

    ______________________________________                                                           Stahylo-                                                     ANTISEPTIC                                                                              Echerichia                                                                              coccus   Streptococcus                                                                           Streptococcus                              TESTED   coli     aureus   group A  group C                                    ______________________________________                                         Septivon-Lavril                                                                         -        ++       -        -                                          Solubacter.sup.R                                                                        -        -        +        -                                          Cyteal   -        +        +        -                                          Hibiscrub                                                                               -        -        -        -                                          Hibisprint                                                                              -        +        +        +                                          Hexomedine.sup.R                                                                        -        -        -        -                                          Foaming paste                                                                           -        -        -        -                                          according to                                                                   the invention                                                                  Cetavlon -        -        -        -                                          ______________________________________                                    

Example 16

Trials similar to those in example 15, but measuring the bacteriostatic activity (capacity to halt the proliferation of the microorganisms) of the antiseptics, were carried out. The foaming paste according to the invention was the same as in Example 15. The following results were obtained (with the same conventions as in Example 15):

    ______________________________________                                                           Stahylo-                                                     ANTISEPTIC                                                                              Echerichia                                                                              coccus   Streptococcus                                                                           Streptococcus                              TESTED   coli     aureus   group A  group C                                    ______________________________________                                         Septivon-Lavril                                                                         -        +        -        -                                          Solubacter.sup.R                                                                        -        -        -        -                                          Cyteal   -        +        +        -                                          Hibiscrub                                                                               -        -        -        -                                          Hibisprint                                                                              -        +        +        -                                          Hexomedine.sup.R                                                                        -        -        -        -                                          Foaming paste                                                                           -        -        -        -                                          according to                                                                   the invention                                                                  Cetavlon -        -        -        -                                          ______________________________________                                    

Example 17

The resistance of Pseudomonas aeruginosa to various reputedly bactericidal active principles or commercial antiseptics was studied in terms of the hardness of the water and the presence or absence of proteins. The same trial was carried out on the foaming paste according to the invention (see Example 15).

Mercryl Lauryle is a foaming solution marketed by Laboratoires LABAZ (Paris, France).

Cresol is the name given to three isomeric phenols, ortho, meta and para, C₇ H₈ O, immediate homologues of phenol, HO--C₆ H₄ --CH₃.

In the following table, R denotes resistant (a growth of colonies was observed), S denotes sensitive (no significant growth was observed), P⁺ means that proteins were added to the solvent and P⁻ means that solvent does not contain proteins.

    __________________________________________________________________________            0.2 ml of innoculum                                                                        0.4 ml of innoculum                                                                        0.6 ml of innoculum                                    contact time 10 mn                                                                         contact time 20 mn                                                                         contact time 30 mn                              ANTISEPTIC                                                                            Distilled                                                                            Hard  Distilled                                                                            Hard  Distilled                                                                            Hard                                      PRODUCTS                                                                              water water water water water water                                     TESTED P+ P- P+ P- P+ P- P+ P- P+ P- P+ P-                                     __________________________________________________________________________     Crude cresol                                                                          S  S  S  S  R  S  S  R  R  R  R  R                                      Mercryl lauryle                                                                       R  S  R  R  R  R  R  R  R  R  R  R                                      Solubacter.sup.R                                                                      S  S  R  S  R  R  R  R  R  R  R  R                                      Foaming paste                                                                  according to                                                                          S  S  S  S  S  S  S  S  S  S  S  S                                      the invention                                                                  Formaldehyde                                                                          S  S  S  S  S  S  S  S  S  S  S  S                                      Hexomedine.sup.R                                                                      S  S  S  S  R  S  R  R  R  R  R  R                                      Hibisprint                                                                            S  S  S  S  S  S  R  R  R  R  R  R                                      Cetavlon                                                                              S  S  R  S  S  S  S  S  S  S  S  S                                      Zinc sulphate                                                                         R  R  R  R  R  R  R  R  R  R  R  R                                      __________________________________________________________________________

The following galenical forms can be (for example, and without limitation) used by way of a dermatological local antiseptic (skin, mucosa, etc.):

FOAMING BAR:

    ______________________________________                                         benzalkonium chloride  2%                                                      LiF                    0.04%                                                   NaF                    1.5%                                                    ______________________________________                                    

Excipients: foaming synthetic base, preservative such as KATHON (registered trademark), purified water, perfume qs. 100%.

FOAMING PASTE: Same formulations as for the foaming bar, except for the excipient formulated as a paste.

MOISTURIZING CREAM:

    ______________________________________                                         benzalkonium chloride  0.2%                                                    LiF                    0.04%                                                   NaF                    0.8%                                                    ______________________________________                                    

Excipients: emulsive agent, moisturizing principle, wheatgerm oil, sweet almond oil, liquid paraffin, preservative such as KATHON (registered trademark), purified water qs. 100%.

IV--APPLICATION OF THE INVENTION IN THE FIELD OF ANTIBIOTIC THERAPY Example 18

The unicellular living organism chosen for the trials is Pseudomonas aeruginosa (wild-type strain originating from hospital sources).

The same methodology as for Examples 11 and 12 was follows. Bacteriostatic analyses were performed with a number of antibiotics belonging to benzylpenicillins, and aminopenicillins, and first and second generation cephalosporins, namely:

a carboxypenicillin: ticarcillin,

three ureidopenicillins: azlocillin, piperacillin, apalcillin,

five cephalosporins: cefoperazone, cefotaxime, ceftriaxone, cefsulodin, ceftazidime.

The determination of the MIC was carried out by the microdilution method in liquid medium on plates. Aqueous solutions of each antibiotic are prepared in MUELLER HINION broth, to obtain dilutions in a geometric ratio of 2. The solvent is sterile double-distilled water.

The preparation of the bacterial inoculum is made with a culture of Pseudomonas aeruginosa, with agitation on a water bath at 37° C. for 4 to 6 hours, giving 10⁶ bacteria/ml.

The bacterial inoculum and the dilution of antibiotic are placed in the plates, and incubated for 18 to 24 hours at 37° C.--the MIC is read in the depressions. Of 958 strains tested, 235 proved resistant to ticarcillin. Of these 235, 69 do not produce a β-lactamase and 166 produce a β-lactamase, namely: 50% a PSE β-lactamase, 17% a TEM β-lactamase, 28% an OXA 8-lactamase and 5% a cephalosporinase.

Three trials were performed per antibiotic and per bacterial inoculum. The trials were performed first with the antibiotics alone (trials a)§, and then with the addition of a dose of 1 mg/l of an aqueous solution of lithium fluoride (trials b)§ to the dilution of each one in every cup of every plate.

The results of the trials are gathered in the following table:

    __________________________________________________________________________                               STRAIN lic R                                                       STRAIN                                                                               STRAIN                                                                               βLac.sup.+                                                    lic S lic S MIC (μg/ml)                                                     βLac.sup.-                                                                      βLac.sup.-                                                                               CEPHALO-                                    TRIAL                                                                              ANTIBIOTIC                                                                               MIC (μg/ml)                                                                       MIC (μg/ml)                                                                       TEM                                                                               PSE                                                                               OXA                                                                               SPORINASE                                   __________________________________________________________________________     1)  TICARCILLIN                                                                              32    512   1024                                                                              4096                                                                              512                                                                               256                                         2a) AZLOCILLIN +                                                                             8     32    64 64 64 128                                         2b) LiF 1 mg/l                                                                               4     32    64 64 64 128                                         3a) PIPERACILLIN +                                                                           8     16    64 128                                                                               64 128                                         3b) LiF       8     8     64 64 64 128                                         4a) APALCILLIN +                                                                             4     16    16 64 32 16                                          4b) LiF       2     8     8  32 32 16                                          5a) CEFOPERAZONE +                                                                           8     16    32 64 64 64                                          5b) LiF       4     16    32 32 64 64                                          6a) CEFOTAXIME +                                                                             16    32    32 32 32 256                                         6b) LiF       8     16    32 16 16 256                                         7a) CEFTRIAXONE +                                                                            16    32    32 32 32 256                                         7b) LiF       8     16    32 16 32 128                                         8a) CEFSULODIN +                                                                             4     16    32 32 16 64                                          8b) LiF       4     16    32 16 16 64                                          9a) CAFTAZIDIME +                                                                            4     4     4  2  4  32                                          9b) LiF       2     4     4  2  4  32                                          __________________________________________________________________________

Abbreviations used:

Tis S : sensitive to ticarcillin

Tic R : resistant to ticarcillin

βLac⁻ : constitutive not producing β-lactamase

βLac⁺ : constitutive producing β-lactamase

MIC : average MIC value

It is thus noted that the addition of lithium fluoride comparison between trials a) and b)§ enables the MIC with respect to the Tic S and Tic R strains of almost all the antibiotics to be decreased.

Example 19

With the same methodology as above, the MIC values of benzalkonium chloride alone, in the presence of the KATHON CG (registered trademark) preservative (1 mg/l), and then in the presence of KATHON CG (registered trademark) (1 mg/l) and lithium fluoride (1 mg/l), were determined. 200 wild-type strains of Pseudomonas aeruginosa were tested.

The following results were observed:

    __________________________________________________________________________              STRAIN Tic                                                                           STRAIN Tic                                                               S βLac.sup.-                                                                    R βLac.sup.+                                                                    STRAIN Tic R βLac.sup.+  MIC (mg/l)                  ANTIBIOTIC                                                                              MIC (mg/l)                                                                           MIC (mg/l)                                                                           TEM                                                                               PSE                                                                               OXA                                                                               CEPHALOSPORINASE                                 __________________________________________________________________________     TICARCILLIN                                                                             32    512   1024                                                                              4096                                                                              512                                                                               256                                              BENZALKONIUM                                                                            32    64    32 64 64 128                                              CHLORIDE                                                                       +KATHON CG.sup.R                                                                        16    32    32 64 32 128                                              (1 mg/l)                                                                       +KATHON CG.sup.R                                                                        16    16    32 32 16 32                                               +(1 mg/l)                                                                      __________________________________________________________________________

A big improvement is hence observed in the presence of lithium fluoride, this being the case even on the strains producing cephalosporinase.

Example 20

The same trial as in Example 19 was performed, but with nonoxinol 9 as the primary active principle: 200 wild-type strains of Pseudomonas aeruginosa were also tested. The results are as follows:

    __________________________________________________________________________              STRAIN Tic                                                                           STRAIN Tic                                                               S βLac.sup.-                                                                    R βLac.sup.+                                                                    STRAIN Tic R βLac.sup.+  MIC (mg/l)                  ANTIBIOTIC                                                                              MIC (mg/l)                                                                           MIC (mg/l)                                                                           TEM                                                                               PSE                                                                               OXA                                                                               CEPHALOSPORINASE                                 __________________________________________________________________________     TICARCILLIN                                                                             32    512   1024                                                                              4096                                                                              512                                                                               256                                              NONOXINOL 9                                                                             64    128   128                                                                               64 128                                                                               256                                              +KATHON CG.sup.R                                                                        32    128   128                                                                               64 128                                                                               256                                              (1 mg/l)                                                                       +KATHON CG.sup.R                                                                        32    64    64 64 64 128                                              +LiF                                                                           (1 mg/l)                                                                       __________________________________________________________________________

Example 21

The activity of CEFOTAXIME was tested, alone and then in the presence of lithium fluoride, with respect to pathogenic strains other than Pseudomonas aeruginosa.

The following table expresses the concentrations (mg/l) of cefotaxime inhibiting 50% of the strains.

    ______________________________________                                                                   50% INHIBITORY                                                                 CONCENTRATION OF                                               50% INHIBITORY  CEFOTAXIME IN THE                                              CONCENTRATION OF                                                                               PRESENCE OF LiF                                      STRAIN    CELFOTAXIME ALONE                                                                              at 1 mg/l                                            ______________________________________                                         Escherichia coli                                                                         0.023           0.01                                                 K12 J53                                                                        Escherichia coli                                                                         0.023           0.023                                                K12 PIP 111                                                                    Escherichia coli                                                                         0.023           0.023                                                K12 PIP 55                                                                     Escherichia coli                                                                         0.18            0.18                                                 SOL                                                                            Klebsiella 1103                                                                          0.023           0.01                                                 Klebsiella U28                                                                           0.28            0.16                                                 Enterobacter T45                                                                         0.05            0.023                                                Enterobacter P49                                                                         27.2            23.2                                                 Proteus morganii                                                                         0.01            0.01                                                 F20                                                                            Serratia 1123                                                                            0.076           0.076                                                Serratia MO1117                                                                          0.30            0.30                                                 ______________________________________                                    

Example 22

The antibiotic activity of amoxicillin (type A penicillin), alone or combined with lithium fluoride, was tested with respect to hospital strains of Haemophilus influenzae producing β-lactamase.

The methodology employed is the same as above. The concentration of amoxicillin alone inhibiting the strains to 100% varied between 32 ppm and 64 ppm.

A concentration of 6 ppm of amoxicillin in the presence of 8 ppm of lithium fluoride enabled the strains to be inhibited to 100% in a time varying between 10 and 12 hours.

By way of comparison, a concentration of amoxicillin of 4 ppm in the presence of 1 ppm of clavulanic acid obtained, for example, by the proprietary pharmaceutical product known as AUGMENTIN and marketed by Laboratories BEECHAM-SEVIGNE (Paris, France) enabled the strains to be inhibited to 100% in 24 hours.

It is hence found that the combination of LiF with amoxicillin enables the threshold of sensitivity of 16 ppm for Haemophilus influenzae to be crossed.

Example 23

The antibiotic activity of amoxicillin, alone or combined with lithium fluoride, was tested with respect to hospital strains normally resistant to amoxicillin (inhibitory concentration greater than 16 ppm).

The results are as follows:

    ______________________________________                                                            100% INHIBITORY                                                                CONCENTRATION OF                                                               AMOXICILLIN IN THE                                                 100% INHIBITORY                                                                            PRESENCE OF LITHIUM                                                CONCENTRATION                                                                              FLUORIDE (LiF)                                                       OF AMOXICELLIN                                                                               Amoxicillin LiF                                         STRAIN   (ppm)         (ppm)       (ppm)                                       ______________________________________                                         Staphylococcus                                                                          32            8           8                                           aureus                                                                         β-Haemolytic                                                                       16            4           8                                           streptococcus                                                                  Klebsiella                                                                              64            2           8                                           pneumoniae                                                                     Haemophilus                                                                             >128          32          8                                           influenzae                                                                     Escherichia                                                                             >128          32          8                                           coli                                                                           (TEM plasmid)                                                                  ______________________________________                                    

Example 24

The antibiotic activity of ampicillin (type A penicillin of the beta-lactam family), alone or in the presence of lithium fluoride, was tested with respect to various strains.

The results are as follows:

    ______________________________________                                                              MINIMAL INHIBITORY                                                             CONCENTRATION OF                                                               OF AMPICILLIN                                                       MINIMAL INHIBI-                                                                           IN THE PRESENCE                                                     TORY CONCEN-                                                                              OF LITHIUM                                                          TRATION OF FLUORIDE (LiF)                                                        AMPICILLIN IN                                                                               Ampicillin                                                                               LiF                                         STRAIN      mg/l         in mg/l   in mg/l                                     ______________________________________                                         Escherichia coli                                                                           4            2         1                                           ATCC 25 922                                                                    Streptococcus faecalis                                                                     8            4         5                                           ATCC 25 212                                                                    Escherichia coli                                                                           2-8          1-4       5                                           (10 wild-type strains)                                                         Streptococcus faecalis                                                                     4-8          2         5                                           (10 wild-type strains)                                                         Group C     32           8         1                                           streptococcus                                                                  Pseudomonas 16-32        4-8       5                                           aeruginosa                                                                     ______________________________________                                    

    ______________________________________                                                                  MINIMAL INHIBITORY                                                             CONCENTRATION                                                                  OF AMPICILLIN                                                     MINIMAL INHIBI-                                                                             IN THE PRESENCE                                                   TORY CONCEN- OF LITHIUM                                            STRAIN      TRATION OF   FLUORIDE (LiF)                                        RESISTANT   AMPICILLIN IN                                                                               Ampicillin                                                                               LiF                                         TO AMPICILLIN                                                                              mg/l         in mg/l   in mg/l                                     ______________________________________                                         Haemophilus >128         32        5                                           influenzae                                                                     (5 wild-type strains)                                                          Staphylococcus aureus                                                                      >128         64        1                                           (10 strains)                                                                   Staphylococcus aureus                                                                      >128         64        6                                           (strains producing                                                             beta-lactamase)                                                                Entrococcus faecium                                                                        >128         64        5                                           Resistant   64           8         5                                           Escherichia coli                                                               ______________________________________                                    

    ______________________________________                                                                  MINIMAL INHIBITORY                                                             CONCENTRATION                                                                  OF AMPICILLIN                                                     MINIMAL INHIBI-                                                                             IN THE PRESENCE                                       MULTIRESISTANT                                                                             TORY CONCEN- OF LITHIUM                                            STRAINS OFTEN                                                                              TRATION OF   FLUORIDE (LiF)                                        PRODUCING   AMPICILLIN IN                                                                               Ampicillin                                                                               LiF                                         BETA-LACTAMASE                                                                             mg/l         in mg/l   in mg/l                                     ______________________________________                                         Pseudomonas 16-32        4-8       0.8                                         aeruginosa                                                                     Escherichia coli                                                                           64           8         0.8                                         Staphylococcus aureus                                                                      >128         32        0.8                                         Group C Streptococcus                                                                      256          64        0.8                                         Haemophilus >128         32        0.8                                         influenzae                                                                     ______________________________________                                    

Example 25

Combating Bronchopulmonary Infections

The same trials as in Examples 22 and 23 were carried out with tetracycline (cycline family), alone or in the presence of lithium fluoride, on wild-type hospital strains responsible for bronchopulmonary infections.

    ______________________________________                                                                100% INHIBITORY                                                                CONCENTRATIONS                                                                 OF TETRACYCLINE                                                                IN THE PRESENCE                                                    100% INHIBITORY                                                                            OF LITHIUM                                                         CONCENTRATION                                                                              FLUORIDE (LiF)                                                       OF TETRACYCLINE                                                                              Tetracycline                                                                             LiF                                       STRAIN       (ppm)         (ppm)     (ppm)                                     ______________________________________                                         Haemophilus influenzae                                                                      4-8           2-4       4                                         Klebsiella pneumoniae                                                                       0.5           0.25      2                                         ______________________________________                                    

Example 26

Combating Skin Infections

The same trials as in Examples 22, 23 and 25 were carried out on wild-type hospital strains responsible for skin infections with, on the one hand polymyxin 8 alone or in the presence of lithium fluoride, and on the other hand erythromycin (macrolide family), alone or in the presence of lithium fluoride.

The results are as follows:

    ______________________________________                                                                100% INHIBITORY CONCEN-                                                        TRATIONS OF POLYMIXIN B                                          100% INHIBITORY                                                                              IN THE PRESENCE OF                                               CONCENTRATION LITHIUM FLUORIDE (Lif)                                           OF POLYMIXIN B                                                                               Polymixin B Lif                                         STRAIN   (ppm)         (ppm)       (ppm)                                       ______________________________________                                         Group C  >128          64          8                                           streptococcus                                                                  Escherichia                                                                             >128          64          2                                           coli                                                                           Staphylococcus                                                                          >128          64          2                                           aureus                                                                         ______________________________________                                    

    ______________________________________                                                                 100% INHIBITORY                                                                CONCENTRATIONS                                                                 OF ERYTHROMYCIN                                                 100% INHIBITORY                                                                               IN THE PRESENCE OF                                              CONCENTRATION  LITHIUM FLUORIDE (Lif)                                          OF ERYTHROMIYCIN                                                                              Erythromycin                                                                               Lif                                        STRAIN   (ppm)          (ppm)       (ppm)                                      ______________________________________                                         Group C  >0.25          0.125       4                                          streptococcus                                                                  Escherichia                                                                             >64            32          8                                          coli                                                                           ______________________________________                                    

The various examples above show that the addition of lithium fluoride with certain antibiotics makes it possible to envisage the oral or parenteral administration of these antibiotics for combating pathogenic organisms which could not be effectively destroyed previously.

The invention hence relates to an antibiotic product that is administrable, in particular, orally, intravenously or, where appropriate, endolymphatically, characterized in that it contains, on the one hand of the order of 10 mg of lithium fluoride in 10 millilitres of injectable solution, and on the other hand an antibiotic, and an antibiotic product that is administrable intravenously, characterized by concentrations compatible with the physiocochemical characters of the components and their possible toxicity.

Example 27

The use of antibiotic agents in chemical antibiotherapy is limited because of the presence of some strains having a resistant phenotype. Those strains would necessitate the use of amounts which cannot be administered to patients because of secondary effects and/or toxicity.

The following trials show that the combination of Fli as activator with an antibiotic agent such as gentamicine changes the phenotype from resistant to sensible, so that the resistant strains may be clinically combated with this combination.

The strains used were wild-type hospital strains having a resistant phenotype as regards gentamicine: pseudomonas aeruginosa, proteus mirabilis, serratia marcescens.

The phenotype of the strains was determined by diffusion in gradient on selective agar. Then, each group of strains having the same phenotype (resistant or sensible) was seeded on inclined agar having a gradient of concentration of the composition.

This methodology is explained by SZYBALSKI and BRYSON in "Selection de mutants resistant aux antibiotiques", L'antibiogramme, Vigot, p. 259-260.

The results are shown in the following table illustrating the MIC of gentamicine (aminoglycoside antibiotic).

x/y means a concentration read between x ppm and y ppm.

    __________________________________________________________________________                  PSEUDOMONAS                                                                             PROTEUS                                                                               SERRATIA                                          STRAIN       AERUGINOSA                                                                              MIRABILIS                                                                             MARCESCENS                                        __________________________________________________________________________     MIC (ppm or mg/l)                                                                           64/128   32/64  32/64                                             alone on the strains before                                                    separation of the phenotypes                                                   MIC (ppm or mg/l)                                                                           32/64    16/32  16/32                                             of gentamicine alone on                                                        SENSIBLE strains                                                               MIC (ppm or mg/l)                                                                           >32      >32    >32                                               of Fli alone on SENSIBLE                                                       strains                                                                        MIC (ppm or mg/l)                                                                           16/32    4/8    8                                                 of gentamicine in presence                                                                  (Fli at 8/16)                                                                           (Fli at 4/8)                                                                          (Fli at 8)                                        of Fli (ppm or mg/l) on                                                        SENSIBLE strains                                                               MIC (ppm or mg/l)                                                                           256/512  64/128 128/256                                           of gentamicine alone on                                                        RESISTANT strains                                                              MIC (ppm or mg/l)                                                                           64       16/32  32                                                of gentamicine in presence                                                                  (Fli at 16)                                                                             (Fli at 8/16)                                                                         (Fli at 16)                                       of Fli (ppm or mg/l) on                                                        RESISTANT strains                                                              __________________________________________________________________________

Therefore, the MIC of gentamicine in presence of Fli on resistant strains is lowered to values similar to the MIC on sensible strains, and a synergistic effect is obtained by the combination of gentamicine anf Fli on sensible strains.

It is possible to clinically administrate the antibiotic even in presence of resistant strains, and with amounts 50% lower in presence of Fli than the amounts needed with gentamicine alone.

This synergistic activation of Fli has also been similarly demonstrated on other families of antibiotic agents.

Example 28

The synergistic effect of the combination of Fli with an antibiotic agent has been shown by the "chess" test which is a known methodology of dilution in liquid medium (Microdilution transportplate technic for determining in vitro synergy of antimicrobial agents"--DOUGHERTY P. F. Antimicrobial Agent Chemotherapy n° 9.225 228, 1977, or "In vitro activity synergism and testing parameters of amikacyn with comparisons to other aminoside antibiotics" KELLY M. T. Antimicrobial Agents Chemotherapy n° 9.440 447, 1976). Concentration series of two antibiotic substances A and B are prepared in the geometric ratio of 2 are combined and the following values are determined:

MIC A = minimal inhibiting concentration of A alone

MIC B = minimal inhibiting concentration of B alone

MIC A/B = minimal inhibiting concentration A in presence of B

MIC B/A = minimal inhibiting concentration B in presence of A ##EQU1##

The MIC of the bacteriostatic effect is read when the percentage of surviving strains is less or equal to 1%.

The combination of the substances A and B is synergistic as regards the bacteriostatic effect when FIC≦0,75. The combination is additive when FIC=1. The combination is insensible when 1<FIC≦2. The combination is antagonistic when FIC>2.

TRIAL 1

Substance A was tetracycline (concentration serie from 0,25 to 128 mg/l)

Substance B was Fli (concentration serie from 0,25 to 16 mg/l)

The strain was staphylococcus aureus (wild-type hospital resistant strain)

It was found that:

MIC A = 32 mg/l

MIC B = 64 mg/l

MIC A/B = 4 mg/l

MIC B/A = 16 mg/l

FIC = 0,375

The combination is thus demonstrated as synergistic.

TRIAL 2

Substance A was gentamicine

Substance B was Fli

The strain was a very resistant proteus mirabilis (wild-type hospital)

MIC A = 64 mg/l

MIC B = 128 mg/l

MIC A/B = 16 mg/l

MIC B/A = 16 mg/l

FIC = 0,375

TRIAL 3

Substance A was gentamicine

Substance B was Fli

The strain was Klebsiella pneumoniae (wild-type hospital strain)

MIC A = 16 mg/l

MIC B = 128 mg/l

MIC A/B = 4 mg/l

MIC B/A = 32 mg/l

FIC = 0,5

The combination is thus demonstrated as synergistic.

TRIAL 4

Substance A was gentamicine

Substance B was Fli

The strain was pseudomonas aeruginosa (wild-type hospital resistant strain)

MIC A = 64 mg/l

MIC B = 128 mg/l

MIC A/B = 16 mg/l

MIC B/A = 16 mg/l

FIC = 0,375

Example 29

It is known that a strain develops a resistance against an antibiotic substance with which it is left in contact, by a phenotypical mutation procedure.

This phenomena has been studied on pseudomonas aeruginosa and Escherichia coli (wild-type hospital strains) which commonly develop resistance against aminosides. The antibiotic substance choosen was thus gentamicine which is an aminoside antibiotic.

And the growth of the resistance was studied with gentamicine alone, then with gentamicine in presence of Fli.

The trial was conducted on 12 days, and the MIC of gentamicine alone and of gentamicine in presence of Fli (4 ppm) was determined each day on transportplates in liquid medium with a serie of dilution in Mueller/Hinton agar.

The MBC (Minimal Bactericide Concentration) was also determined from the solution after determination of the MIC seeded on agar adapted to the strain. The MBC is read for a percentage of survivors less than 0,20%.

It appears that no resistance is developped by the strain against Fli, but a resistance corresponding to 8 to 16 times the initial MIC is developped by gentamicine alone, the third or fourth day.

In presence of gentamicine with Fli (4 ppm) the resistance appears later) the fourth or sixth day) an only corresponds to 4 to 8 times the initial MIC.

Therefore the presence of Fli retards the creation of the resistance and reduces to the half the capability of each strain for developping such resistances.

Example 30

The kinetics of the bacterial growth of serratia marcescens and salmonella spirochete has been studied with the "Time killing curve" methodology, for gentamicine (64 ppm) alone, Fli (32 ppm) and gentamicine (64 ppm) in presence of Fli (32 ppm). The bacterial growth is determined by spectrophotometric reading of the optical density for a wavelength of 600 nm.

The following table illustrates the optical density (1/100) obtained:

    __________________________________________________________________________     STRAIN               0 h 00                                                                             3 h 00                                                                             6 h 00                                                                             24 h 00                                       __________________________________________________________________________     SERRATIA  INOCULUM    8, 5                                                                              --  --  --                                            MARCESCENS                                                                               GROWTH SAMPLE                                                                             --  36  45  49                                                      GENTAMICINE                                                                               --   5, 5                                                                               5   4                                                      Fli        --  13  15  18                                                      GENTAMICINE + Fli                                                                         --   2, 5                                                                               1, 5                                                                               0, 7                                         SALMONELA INOCULUM   11  --  --  --                                                      GROWTH SAMPLE                                                                             --  29  37  44                                                      GENTAMICINE                                                                               --   8   7   5, 5                                                   FLi        --  14  14, 5                                                                              20                                                      GENTAMICINE + Fli                                                                         --   5   3   1, 5                                         __________________________________________________________________________

It is thus shown a synergistic activity of gentamicine combined with Fli.

Example 31

The MIC of tetracycline was studied on Klebsiella pneumoniae (wild-type hospital strain).

The trial was conducted with tetracycline alone, tetracycline in presence of FNa and tetracycline in presence of Fli.

    ______________________________________                                               TETRACYCLINE TETRACYCLINE TETRACYCLINE                                         alone        + FNa (8 ppm)                                                                               + Fli (8 ppm)                                  ______________________________________                                         MIC   512          256/512      128                                            (ppm)                                                                          ______________________________________                                    

It is thus demonstrated a better unexpected activation of tetracycline by Fli than by FNa.

Example 32

The MIC of doxycycline was studied on pseudomonas aeruginosa (wild-type hospital strain) in the same conditions as for example 31.

    ______________________________________                                               DOXYCYCLINE  DOXYCYCLINE  DOXYCYCLINE                                          alone        + FNa (8 ppm)                                                                               + Fli (8 ppm)                                  ______________________________________                                         MIC   1000         500/1000     256/500                                        (ppm)                                                                          ______________________________________                                    

Thus Fli is a surprisingly better activator of doxycycline than Fna.

Example 33

The efficiency in vivo of the administration of doxycycline in presence of Fli has been measured and compared with the efficiency of the administration of doxycycline alone.

The antibiotic composition (doxycycline alone or with Fli) has been administered to a healthy patient, and the antibiotic activity of the patient's blood has been determined before the first administration, the day of the administration, and three days later. The delay of three days is necessary because of the low solubility of Fli: the pertinent concentration of Fli is only obtained after the end of the second day after its administration.

The patient had taken the antibiotic during the three days, 2 capsules of 250 mg each day at 8 a.m. A blood sample was taken 3 hours after each administration.

And the antibiotic activity was twice determined by a microbiological titration using the inhibition holes methodology on a standard strain ATCC 25922 of Escherichia coli.

Holes of 6 nm diameter were made on a Mueller Hinton agar and 100 μl of plasma were injected in each hole. The agar was then seeded with an inoculum of Escherichia coli of 10 UFC/ml.

After an incubation of 24 hours at 37° C., the diameters around each holes (in which the strain was killed) were read.

In the first trial, the capsules of 250 mg were constituted of 110 mg of doxycycline chlorhydrate and of lactose.

The inhibition diameters were measured as follows

J0 = the day before the first administration

J1 = the day of the first administration

J3 = the third day of administration

    ______________________________________                                                         INHIBITION DIAMETER (mm)                                       DAY OF MEASURE  DOXYCYCLINE ALONE                                              ______________________________________                                         J 0             7                                                              J 1             8-9                                                            J 3             11-12                                                          ______________________________________                                    

The second trial was conducted similarly with gelules of 250 mg constituted of 110 mg of doxycycline, 53 mg of Fli and lactose.

    ______________________________________                                                         INHIBITION DIAMETER (mm)                                       DAY OF MEASURE  DOXYCYCLINE + Fli                                              ______________________________________                                         J 0             7                                                              J 1             10-11                                                          J 3             14-15                                                          ______________________________________                                    

Therefore the better antibiotic activity of the activated antibiotic is demonstrated in vivo.

Example 34

Clinical experiments on ill patients were conducted with antibiotic compositions activated with Fli.

The antibiotic agent dose in 250 mg capsules was lowered to 60% of the normally administered dose (defined by VIDAL DICTIONARY, France, 1987), and an appropriate dose of Fli was added. The activated capsules were administered with the same methodology as for the non-activated antibiotic capsules (as indicated in the VIDAL DICTIONARY).

The appropriate dose of Fli was defined to have an amount of lithium in the blood of 1,5 ppm after 2 hours. Thus, when only one capsule is administered per day (for example for doxyclycline) an approximative dose of 50 mg of Fli is used in the capsule. When two capsules are administered per day (for example for bactrim (registered trademark) the dose of Fli per capsule is about 25 mg.

47 experiments were conducted on patients who had caught pneumopathies such as bronchitis, or cystisis due to staphylococci. Various kinds of antibiotic agents were used: doxycycline, amoxicilline, spiramycine, tetracycline, josamycine, bactrim (registered trademark) (sulfamide+diaminopyrimidine) . . .

No case of intolerance was observed.

Only one check was met with a 65 years old patient attached by bronchitis treated with activated spiramycine. All the other patients were cured in 4 to 6 days of treatment, though the antibiotic dose was only 60% the normally prescribed dose.

Furthermore some of the patients thus cured had been attacked by chronicle affections or had been previously treated by a firts standard antibiotic treatment without success.

Therefore the surprising efficiency of the activation by Fli has been clinically demonstrated in vivo.

Example 35

Strains Gram+ and Gram- (wild-type hospital) having a resistant phenotype vis a vis standard firstly and secondly used antibiotherapy were experimented on activated antibiotics.

250 experiments were conducted on the following strains: Streptococcus group B, C, D, F, G, Staphylococcus aureus, Staphylococcus epidermitis, Klebsiella pneumoniae, Klebsiella spirochete, Klebsiella oxytoca, Proteus mirabilis, Serratia marcescens, Escherichia coli, Citrobacter freundii, Enterobacter cloacae, Enterobacter calcoaceticus, Salmonella spirochete, Salmonella typhimurium, Pseudomonas aeruginosa, Haemophilus influenzae, Haemophilus sphrophilus, Acinetobacter calcoaeticus.

The antibiotic agents used were (depending on each strain, the 10 to 20 antibiotic agents commonly used on the strains were experimented without Fli and in presence of Fli at 4 ppm): Penicilline (group G), Ampicilline, Erythromicine, Tetracycline, Amoxicilline, Doxycycline, Minocycline, Trimethoprime, Josamycine, Streptomycine, Gentamicine, Cefazoline, Chloramphenicol, Framycetine, Bactrim (registered trademark), Thiamphenicol, Pristinamycine, Lincomycine, Troleandomycine, Oxacilline, Cefalotine, Mezlocilline, Cefalexine, Cefotaxime, Amoxicilline+clavulanic acid, Lymecycline, Netilmicine, Nalidixic acid, Pefloxacine, Tobramycine, Amikacine, Norfloxacine, Cefsulodine, Polymixine B, Cefoperazone, Kanamycine, Midecamycine, Nitro-furantoine, Cotrimoxazole, Ticarcilline, Piperacilline, Ofloxacine, Fosfomycine, Ticarcilline+clavulanic acid. Therefore all the families of antibiotic agents were experimented.

The results of the experiments have been given in term of phenotype: resistant when the MIC is superior to a given threshold value (depending on the antibiotic agent, commonly about 16 ppm), insensible when the MIC is around the threshold value, sensible when the MIC is inferior to the threshold value.

In the case of streptococcus group F with penicilline group G, the insensible phenotype was not changed by the presence of Fli (4 ppm).

In the case of Klebsiella pneumonia with cefoperazone and staphylococcus aeurus with amoxicilline or erythromycine the resistant phenotype was not changed by the presence of Fli (4 ppm).

In all the other cases, the phenotype changed from resistant or insensible to insensible of sensible.

In 145 experiments, the strain became totally sensible vis a vis the activated antibiotic. And 73 totally resistant strains were even changed to totally sensible with the activation by Fli. The other totally resistant strains representing 76 experiments were changed to insensible strains.

It is thus possible with the invention to combat some resistant pathogen microorganism, where this was not possible with classical antibiotherapy. The mere activation of antibiotics with 4 ppm of Fli, which is a totally innocuous dose, surprisingly allows the curation of high resistant strains such as haemophilus influenzae, Klebsiella pneumoniae or others.

It is noted that an "antibiotic" agent may be described as a germicidal agent which can be introduced into the human or animal body at such concentrations that it can maintain its efficacy without any resultant toxicity or side effects. 

What is claimed is:
 1. A method for inhibiting or destroying unicellular organisms which comprises administering a composition which comprises an active ingredient, anda synergistically effective amount of a potentiator which is lithium fluoride, said amount being effective to reduce the amount of said active ingredient necessary to inhibit or destroy said unicellular organisms, said amount also being at least 0.2% by weight but less than the amount of said active ingredient.
 2. The method for inhibiting or destroying unicellular organisms of claim 1, wherein the unicellular organisms are selected from the group consisting of protozoa, microbes, bacteria, fungi and yeasts.
 3. The method for inhibiting or destroying unicellular organisms of claim 1, wherein the active ingredient is selected from the group consisting of betalactamins, macrolides, polypeptidic antibiotics, phenicolated antibiotics, rifamycins, lincosanides, streptogramines, sulfamides, trimethoprimes, aminocides, cyclins, quinolones and their derivatives and combinations thereof.
 4. A composition for inhibiting or destroying unicellular organisms which comprises an active ingredient, anda synergistically effective amount of a potentiator which is lithium fluoride, said amount being effective to reduce the amount of said active ingredient necessary to inhibit or destroy said unicellular organisms, said amount also being at least 0.2% by weight but less than the amount of said active ingredient.
 5. The composition for inhibiting or destroying unicellular organisms of claim 4, wherein the unicellular organisms are selected from the group consisting of protozoa, microbes, bacteria, fungi and yeast.
 6. The composition for inhibiting or destroying unicellular organisms of claim 4, wherein the active ingredient is selected from the group consisting of betalactamins, macrolides, polypeptidic antibiotics, phenicolated antibiotics, rifamycins, lincosanides, streptogramines, sulfamides, trimethoprimes, aminocides, cyclins, quinolones and their derivatives and combinations thereof. 